Comparative Pollen Spectra of Tetragonisca Angustula (Apidae, Meliponini) from the Lower Amazon (N Brazil) and Caatinga (NE Brazil) Jaílson S

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Comparative pollen spectra of Tetragonisca angustula (Apidae, Meliponini) from the Lower Amazon (N Brazil) and caatinga (NE Brazil) Jaílson S. de Novais, Ana Cristina A. Garcêz, Maria Lúcia Absy, Francisco de Assis R. dos Santos

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Jaílson S. de Novais, Ana Cristina A. Garcêz, Maria Lúcia Absy, Francisco de Assis R. dos Santos. Comparative pollen spectra of Tetragonisca angustula (Apidae, Meliponini) from the Lower Ama- zon (N Brazil) and caatinga (NE Brazil). Apidologie, Springer Verlag, 2015, 46 (4), pp.417-431. 10.1007/s13592-014-0332-z. hal-01284456

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HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Apidologie (2015) 46:417–431 Original article * INRA, DIB and Springer-Verlag France, 2014 DOI: 10.1007/s13592-014-0332-z

Comparative pollen spectra of Tetragonisca angustula (Apidae, Meliponini) from the Lower Amazon (N Brazil) and caatinga (NE Brazil)

1,2 1 2 Jaílson S. de NOVAIS , Ana Cristina A. GARCÊZ , Maria Lúcia ABSY , 3 Francisco de Assis R. dos SANTOS

1Centro de Formação Interdisciplinar and Laboratório de Botânica Taxonômica, Universidade Federal do Oeste do Pará, Rua Vera Paz, s/n, Salé, 68035-110, Santarém, Pará, Brazil 2Laboratório de Palinologia, Coordenação de Biodiversidade, Instituto Nacional de Pesquisas da Amazônia, Av. André Araújo, 2.936, Petrópolis, 69067-375, Manaus, Amazonas, Brazil 3Laboratório de Micromorfologia Vegetal, Departamento de Ciências Biológicas, Universidade Estadual de Feira de Santana, Av. Transnordestina, s/n, 44036-900, Feira de Santana, Bahia, Brazil

Received 18 January 2014 – Revised 26 September 2014 – Accepted 21 October 2014

Abstract – Tetragonisca angustula is a stingless bee species widely distributed in Brazil. Past studies have shown a rich native flora supplying this meliponine with pollen and nectar. The aim of this study was to compare the food provisions of T. angustula in two areas in the Lower Amazon mesoregion in the state of Pará and two areas of caatinga vegetation in the state of Bahia, Brazil. We used a published data set of 57 samples of honey and 46 samples of pollen stored in colonies of T. angustula . The honeys from the Lower Amazon and from the caatinga displayed 23 and 16 pollen types with frequency higher than >10 %, respectively, with emphasis on the families Fabaceae, Solanaceae, and Urticaceae. In the pollen stored samples, 18 and 11 pollen types were found, respectively. Cluster analyses (Dice similarity) performed with these published pollen spectra of the honeys grouped the samples from each geographic region with higher similarity than the analyses performed with the pollen spectra of the stored pollen. cluster analysis / Dice similarity / entomopalynology / honey / melissopalynology / pollen stored / stingless bees

1. INTRODUCTION distributed in time and space (Fisch et al. 1998). In contrast, the seasonally dry forests of the The Amazon Forest and the caatinga (dry for- caatinga encompass different mosaics of vegeta- est) together form more than 5 million km2, equiv- tion subjected to a rigorous precipitation deficit alent to approximately 60 % of the Brazilian terri- (Prado 2003). Disturbances in both of these biomes tory (IBGE 2004). The Amazon Forest, extending have threatened native bee populations and hin- over eight Latin-American countries in addition to dered the environmental services provided by them Brazil, is the largest tropical rainforest of the planet, (Freitas et al. 2009; Maia-Silva et al. 2012). with a high precipitation level irregularly Recently, in independent palynological studies, we investigated the pollen spectra of honey and Electronic supplementary material The online version of pollen stored by Tetragonisca angustula (Latreille, this article (doi:10.1007/s13592-014-0332-z) contains 1811) (Apidae: Meliponini) from Amazonian supplementary material, which is available to authorized (Novais and Absy 2013; Novais and Absy in users. press)andcaatinga (Novais et al. 2013, 2014) Corresponding author: J. Novais, areas. [email protected] In this manuscript, we performed a compara- Manuscript editor: James Nieh tive palynological analysis of the food provisions 418 J.S. de Novais et al. of T. angustula basedonthepublisheddatamen- honey and pollen, respectively) and Ruy Barbosa (14 tioned above. The data on the pollen types found and 13 samples)] (Novais and Absy 2013; Novais et al. has been previously published but not qualitative- in press; Novais and Absy 2013, 2014). ly compared. With this study, we compiled the The protocol for the collection of material, the data set of plants foraged by T. angustula in two laboratory procedures adopted, and the qualitative biomes (Amazon and caatinga ) and also perform and quantitative palynological analysis performed a cluster analysis aiming to better understand the were previously described by Novais and Absy similarity of pollen diet of this species in different (2013) and Novais and Absy (2013). The technique regions in Brazil, one wet and another dry. of acetolysis (Erdtman 1960) was used to process all the samples. For the purpose of this study, a sample 2. MATERIALS AND METHODS consists of 5 mL of honey or 2–3gofstoredpollen, collected from one colony of T. angustula in each 2.1. Study sites study site. After chemical processing, four glass slides were prepared and at least 500 pollen grains We used data obtained from four study sites: two per sample were identified and quantified. From located in the Lower Amazon mesoregion (Pará State, these data, the following frequency classes were NBrazil)andtwoareasinthecaatinga (Bahia State, established, as proposed by Louveaux et al. (1978): NE Brazil) (Figure 1). Further information about these predominant pollen (>45 % of the grains counted in – areas can be found in Novais and Absy (2013) and a sample), secondary pollen (16 45 %), important – Novais et al. (2013, 2014). minor pollen (3 15 %), and minor pollen (<3 %).

2.2. Collection, laboratory processing, and 2.3. Comparative analysis palynological analysis Comparative analysis was performed with the pollen In total, 103 samples were studied: 57 samples of spectra obtained, considering only the pollen types with honey and 46 samples of pollen stored by T. angustula . frequencies higher than 10 % in at least one of the The samples were collected in between September 2010 samples analyzed because these types represent the and January 2012 in the study sites of the Lower most significant floral resources for the bees (Ramalho Amazon [Belterra (16 and 15 samples of honey and et al. 1985). We have not included in this analysis pollen pollen, respectively) and Santarém (14 and 8 samples)] types which frequencies were lower than 10 % in all and in the caatinga [Itaberaba (13 and 10 samples of samples.

Figure 1. Location of the study sites in the Lower Amazon (map to the left ) and in the caatinga (map to the right ). In the map of the South America, in the center, Brazil is colored in gray, with emphasis on the states of Pará, in the north region, and Bahia, in the northeast region. The municipalities where the study was performed are represented by letters : A Santarém, B Belterra, C Ruy Barbosa, D Itaberaba. Melissopalynology of Amazon and caatinga 419

We performed a cluster analysis, using the Dice In total, the honeys from the Amazonian study similarity coefficient (McCune and Grace 2002), sites displayed 23 representative pollen types, to evaluate the formation of groups among the representing 15 botanical families. In decreasing samples studied based on the pollen types identi- order, Warszewiczia coccinea , Microtea , fied in the samples. For the honeys, the 57 sam- Byrsonima , Cecropia ,andBorreria verticillata ples and the 36 pollen types identified were con- occurred in more than 50 % of the samples sidered for the presence–absence data matrix (Figure 2a). In the honeys from the caatinga ,16 (Hammer et al. 2001). Similarly, for the stored pollen types and 11 botanical families were re- pollen, the 46 samples and the 29 pollen types corded, from which Prosopis juliflora , Solanum , identified were used for the data matrix. Heteropterys ,andSchinus were the most com- Therefore, we evaluated the presence of structured monly observed pollen types (Figure 2b). patterns, considering the period of collection and The pollen stored samples from the the different sampling areas. This analysis consid- Amazonian study sites, in contrast, displayed ered only the presence of the pollen types in the 18 representative pollen types, representing individual samples, disregarding the absence of a 17 botanical families. The pollen types given pollen type as an indication of similarity. Cecropia and B. verticillata occurred in The similarity analysis was performed with the more than 50 % of the samples (Figure 3a). software PAST—PAlaeontological STatistics , ver- The pollen stored samples from the caatinga sion 2.16 (Hammer et al. 2001). The climatic data revealed 11 pollen types and seven botanical of the study sites were obtained from the families. Of these types, the most frequent Brazilian National Institute of Meteorology were P. juliflora , Solanum , Senna (Instituto Nacional de Meteorologia, INMET macranthera , Brosimum ,andHeteropterys 2013), except for the municipality of Ruy (Figure 3b). Barbosa, Bahia, where there is no meteorological The honey samples from the Lower Amazon station to record such data. A given month was displayed eight pollen types classified as “pre- considered dry when the total monthly precipita- dominant pollen”: B. verticillata , Byrsonima , tion, in millimeters, was equal to or lower than Cecropia , Chamaecrista , Clidemia hirta , double the mean monthly temperature, expressed Diodia , Solanum ,andW. coccinea (Table I). in degrees Celsius (P ≤2T ) (Bagnouls and In contrast, the honeys from the caatinga re- Gaussen 1962). vealed five predominant pollen types: Cecropia , Gomphrena demissa , P. juliflora , Schinus ,and 3. RESULTS Solanum (Table I). Furthermore, we emphasize that the pollen types Cecropia and Solanum were 3.1. General considerations of the pollen part, as predominant pollen types, of the foraging spectra and the most representative pollen behavior of T. angustula in both the Lower types Amazon and the Bahian caatinga (Table I and Figure 2). The honey samples revealed a higher number The pollen stored samples of T. angustula from of representative pollen types (those with a fre- the Lower Amazon revealed the following pre- quency higher than 10 % in at least one sample) dominant pollen types: Byrsonima , Cecropia , than the pollen stored samples. The honeys from C. hirta , Davilla kunthii , Eriope , Myrcia ,and Santarém, PA (17 pollen types) recorded the Vismia guianensis (Table II). highest number of pollen types, followed by Ruy The pollen stored samples from the caatinga Barbosa, BA (14), Belterra, PA (12), and revealed seven predominant pollen types: Itaberaba, BA (10) (Table I). For the pollen stored Brosimum , Caesalpinioideae type, P. juliflora , samples, Belterra, PA (11 pollen types) stood out Schinus , S. macranthera , Solanum ,and among the municipalities, followed by Santarém, Waltheria (Table II). No predominant pollen type PA (10), Itaberaba, BA (9), and Ruy Barbosa, BA occurred simultaneously in the pollen stored sam- (7) (Table II). ples from both of the biomes. 420 Table I. Pollen spectra indicative, based on the frequency class, of the number of samples in which the most representative pollen types occurred in honey samples of Tetragonisca angustula (Apidae, Meliponini) in the Lower Amazon, state of Pará (PA), north region, and in the caatinga , state of Bahia (BA), northeast region, Brazil.

Plant family Pollen type Belterra—PA Santarém—PA Itaberaba—BA Ruy Barbosa—BA

p s im m = p s im m = p s im m = p s im m =

Amaranthaceae Alternanthera ––41014 –– – – – ––– – – ––– – – Gomphrena demissa ––– – – –– – – – ––– – – 1 –– 3 4 Anacardiaceae Anacardium occidentale – 13 4 8 –– – – – ––– – – ––– – – Tapirira guianensis – 11 4 6 – 2 – 5 7 ––– – – ––– – – Schinus ––– – – –– – – – 211 – 4 347 1 15 Arecaceae Syagrus coronata ––– – – –– – – – – 1 – 3 4 ––189 Asteraceae Emilia fosbergii – 14 5 10 –– – – – ––– – – ––– – – ..d oase al. et Novais de J.S. Mikania ––– – – –– – – – ––189 ––– – – Bignoniaceae Tabebuia ––– – – –– 235 ––– – – ––– – – Bignoniaceae type ––– – – –– 1 – 1 ––– – – ––– – – Burseraceae Protium ––– – – – 1247 ––– – – ––– – – Euphorbiaceae Acalypha ––– – – –– – – – ––– – – ––134 Alchornea ––– – – –– – – – ––213 ––– – – Euphorbia ––– – – –– – – – ––– – – – 12 1 4 Fabaceae Caesalpinioideae type ––– – – –– – – – ––– – – – 1 – 4 5 Chamaecrista 1 – 225 –– – – – ––– – – ––– – – Clathrothropis nitida ––– – – – 12– 3 ––– – – ––– – – Fabaceae type ––– – – –– 2 – 2 ––– – – ––– – – Mimosa tenuiflora ––– – – –– – – – ––– – – ––279 Prosopis juliflora ––– – – –– – – – 163 3 13 158 1 15 Senna ––– – – – 1225 ––– – – ––– – – Senna macranthera ––– – – –– – – – ––– – – – 12 8 11 Lacistemataceae Lacistema ––– – – –– 123 ––– – – ––– – – Lamiaceae Eriope – 22 7 11 –– – – – ––– – – ––– – – Malpighiaceae Byrsonima 333 1 10 11 4 2 8 ––– – – ––– – – Table I (continued)

Plant family Pollen type Belterra—PA Santarém—PA Itaberaba—BA Ruy Barbosa—BA

p s im m = p s im m = p s im m = p s im m =

Heteropterys ––– – – –– – – – – 35 5 13 – 36 3 12 Melastomataceae Clidemia hirta ––– – – 35 1 1 10 ––– – – ––– – – Moraceae Brosimum ––– – – –– – – – ––– – – – 22 2 6 Myrtaceae Myrcia ––– – – – 2 – 6 8 – 13 9 13 ––– – – Syzygium malaccense ––– – – –– 112 ––– – – ––– – – eisplnlg fAao and Amazon of Melissopalynology Phytolaccaceae Microtea – 35 6 14 – 1348 ––– – – ––– – – Rubiaceae Borreria verticillata 167 1 15 –– – – – ––– – – ––– – – Diodia 1 –– 1 2 –– – – – ––– – – ––– – – Warszewiczia coccinea – 32 6 11 12 8 3 14 ––– – – ––– – – Solanaceae Solanum ––– – – 11 2 2 6 272 2 13 174 1 13 Urticaceae Cecropia 1 – 157 13 1 5 10 1 – 1 – 2 ––– – – Total number of pollen types 5 8 11 12 12 511141317 468 7 10 4 8 10 12 14

Only the pollen types with a frequency equal to or higher than 10 % in at least one of the samples analyzed were considered representative. Frequency classes: p , predominant pollen; s , secondary pollen; im , important minor pollen; m , minor pollen; = , total number of samples in which each pollen type was recorded, per locality. Data based on Novais et al. (2013)andNovais

andAbsy(in press) caatinga 421 422 Table II. Pollen spectra indicative, based on the frequency class, of the number of samples in which the most representative pollen types occurred in samples of pollen stored by Tetragonisca angustula (Apidae, Meliponini) in the Lower Amazon, state of Pará (PA), north region, and in the caatinga , state of Bahia (BA), northeast region, Brazil.

Plant family Pollen type Belterra—PA Santarém—PA Itaberaba—BA Ruy Barbosa—BA

p s im m = p s im m = p s im m = p s im m =

Achariaceae Lindackeria paludosa – 1 ––1 ––– – – ––– – – ––– – – Amaranthaceae Alternanthera – 1168 ––– – – ––– – – ––– – – Anacardiaceae Tapirira guianensis – 1157 ––– – – ––– – – ––– – – Schinus –– – – – ––– – – ––– – – 123 5 11 Asteraceae Emilia fosbergii –– – – – ––123 ––– – – ––– – – Araliaceae Schefflera morototoni – 3148 ––– – – ––– – – ––– – – Bignoniaceae Tabebuia – 2349 ––– – – ––– – – ––– – – al. et Novais de J.S. Dilleniaceae Davilla kunthii –– – – – 1 –– 1 2 ––– – – ––– – – Euphorbiaceae Acalypha – 1 – 3 4 ––1 – 1 ––– – – ––– – – Fabaceae Caesalpinioideae type –– – – – ––– – – 1 –– – 1 ––– – – Machaerium –– – – – ––1 – 1 ––– – – ––– – – Prosopis juliflora –– – – – ––– – – 134 2 10 922 – 13 Senna –– – – – ––1 – 1 ––– – – ––– – – Senna macranthera –– – – – ––– – – 1 – 258 ––6410 Zornia echinocarpa –– – – – ––– – – ––– – – ––112 Hypericaceae Vismia guianensis –– – – – 11––2 ––– – – ––– – – Lamiaceae Eriope 1 – 225 ––– – – ––– – – ––– – – Malpighiaceae Byrsonima 43 ––7 1 –– 1 2 ––– – – ––– – – Heteropterys –– – – – ––– – – – 12 2 5 – 32 3 8 Malvaceae Quararibea –– – – – ––– – – ––112 ––– – – Waltheria –– – – – ––– – – 111 – 3 ––– – – Melastomataceae Clidema hirta –– – – – 3 –– – 3 ––– – – ––– – – Moraceae Brosimum –– – – – ––– – – 311 1 6 – 16 4 11 Brosimum paraense – 1 ––1 ––– – – ––– – – ––– – – Melissopalynology of Amazon and caatinga 423 , s – – – 12 7 – 3.2. Pollen groups based on the cluster )and analysis 2013 BA

— The groups obtained by the cluster analysis of the pollen spectra of the honey samples were more homogeneous than those ob- , predominant pollen;

p tained with the pollen spectra of the pollen ––– – ––– – ––– – 215 4 357 6 ––– – stored samples (Figures 4 and 5). sses: The first phenogram obtained (cophenetic co- 2 – – 9 9 – efficient 0.94, Figure 4) initially formed two dis-

1 tinct groups, gathering all the samples from the Lower Amazon in group A and all samples from BA Ruy Barbosa

– the caatinga in group B. However, the level of — similarity of group Awas lower than that of group 1 B(Figure4). – ––– – ––– – 231 3 667 7 ––– – The cluster analysis also revealed the formation of subgroups within the wider groups (A and B), – 2 – – 10 4 gathering the samples per municipality (Figure 4). In crescent order of similarity, the following sub- 1 groups were formed: C (gathering the samples

PA Itaberaba from Belterra), D (Santarém), E (Ruy Barbosa), 12

— and F (Itaberaba), the last with a level of similarity –– – higher than 0.70 (Figure 4). In group A, the predominant and/or secondary ––– – 1 ––– – ––– – 615 5 1 pollen types shared by subgroups C and D were Byrsonima , Cecropia , Microtea , Tapirira – – 12 – 11 10 guianensis ,andW. coccinea (Figure 4). Of these types, only T. guianensis was not among the most frequent pollen types in the honey samples from the Lower Amazon (Figure 4). The pollen types PA Santarém

— Heteropterys , P. juliflora , Schinus ,andSolanum , total number of samples in which each pollen type was recorded, per locality. Data based on Novais and Absy ( 444 were shared as predominant and/or secondary = pollen types in the samples gathered in group B, psimm=psimm=psimm=psimm= –– – – –– – – – –– – – 62 1 1 thus characterizing this cluster (Figure 4). These same pollen types were among those with the

, minor pollen; highest occurrence in the honey samples from

m the caatinga (Figure 2). The second phenogram (cophenetic coefficient 0.96, Figure 5), obtained from the cluster analysis with the data from the pollen stored samples, shows less-consistent groups than those obtained from the cluster analysis with Virola Myrcia Borreria verticillata Solanum Cecropia Total number of pollen types 3 10 7 8 the data from the honey samples (Figure 4). , important minor pollen;

) Two main groups can be observed: group A, im gathering the samples from the Lower 2014 Amazon; and group B, gathering the samples (continued) from the caatinga (Figure 5). However, these groups showed a low similarity level (<0.4), Myristicaceae Myrtaceae Rubiaceae Table II Only the pollen types with a frequency equal to or higher than 10 % in at least one of the samples analyzed were considered representative. Frequency cla Solanaceae secondary pollen; Novais et al. ( Urticaceae

Plant family Pollen type Belterra especially group A (<0.1) (Figure 5). 424 J.S. de Novais et al.

Figure 2. Occurrence (%) of pollen types in 57 samples of honey produced by Tetragonisca angustula (Apidae, Meliponini) in Amazonian areas (a ) and semiarid areas (b ) of Brazil. Only the pollen types with a frequency equal to or higher than 10 % in at least one of the samples analyzed are represented in this diagram.

From these two main groups (A and B), two pollen (Figure 5). Within subgroup E, cluster K additional sub-groups were formed (Figure 5); stood out for gathering 10 samples exclusively however, these sub-groups did not show a signifi- from Ruy Barbosa at a similarity level higher than cant geographic discrimination, in contrast to the 0.7. The predominant pollen types in this cluster first phenogram (Figure 4). In crescent order of (K) were P. juliflora , Schinus ,andSolanum similarity level, the subgroups C and D were gen- (Figure 5). Clusters L and M gathered samples erated for the Lower Amazon, and the subgroups E from both Itaberaba and Ruy Barbosa, with em- and F were generated for the caatinga (Figure 5). phasis on the pollen type Brosimum (Figure 5). Subgroup C gathered, with low similarity (<0.3), The predominant and/or secondary pollen only three pollen stored samples from Santarém, in types shared by more than one subgroup of which the predominant pollen types were C. hirta group A were Byrsonima (G and H), and D. kunthii . Subgroup D gathered samples Cecropia (G, H, and I), and C. hirta (C and from Belterra and Santarém. Within this subgroup, G) (Figure 5). Of these types, Cecropia was smaller clusters were generated, gathering, for ex- among the most frequent pollen types in the ample, samples exclusive to Santarém (clusters G pollen stored samples from the Lower and J) and to Belterra (H and I). Subgroup E Amazon (Figure 3). In contrast, Brosimum (K, gathered pollen stored samples from the two L, and M), Heteropterys (K and M), caatinga areas, whereas subgroup F gathered only P. juliflora (K, L, and M), Schinus (K and the sample collected in Itaberaba in January 2011, M), and Solanum (K,L,andM)werethe with Caesalpinioideae type as the predominant pollen types most shared by subgroups of group Melissopalynology of Amazon and caatinga 425

Figure 3. Occurrence (%) of pollen types in 46 samples of pollen stored by Tetragonisca angustula (Apidae, Meliponini) in Amazonian areas (a ) and semiarid areas (b ) of Brazil. Only the pollen types with a frequency equal to or higher than 10 % in at least one of the samples analyzed are represented in this diagram.

B (Figure 5). Of these, only Schinus was not morototoni (Araliaceae), and Tabebuia among the pollen types with the highest occur- (Bignoniaceae) (Online Resource 2). rence in the pollen stored by T. angustula in the caatinga (Figure 3). 4. DISCUSSION AND CONCLUSION Concerning the climatic data from the study sites, we noted that in Belterra, five months were Despite the high number of pollen types iden- considered dry (September and November 2010; tified in both the honey and pollen stored samples, August, September, and December 2011) a few pollen types effectively contributed to the (Figure 6). In Santarém, only October 2010 and composition of the majority of the pollen spectra September 2011 were dry. In contrast, in obtained. This finding corroborates previous pal- Itaberaba, all the months of 2011 were dry except ynological studies performed in areas of the November (Figure 6). In the honey samples from Central Amazon involving species of Santarém, the cluster formed by the samples col- Frieseomelitta , Melipona , Scaptotrigona ,and lected between February and April 2011 repre- Trigona (Marques-Souza et al. 1996, 2002; sents the peak of precipitation in this region Marques-Souza 2010). In areas of caatinga in (Figures 4 and 6) and the period of dominance the northeast region of Brazil, a similar pattern of the pollen type C. hirta (Online Resource 1). In was also observed in palynological works with contrast, the pollen stored samples from Belterra species of Melipona , Scaptotrigona ,and in the months of September and October 2010, in Tetragonisca (Alves et al. 2006; Novais et al. addition to those of June, August, and September 2006; Santana et al. 2011). 2011, formed a cluster representing the months In general, several pollen types were recorded that were dry or had low precipitation in both the pollen stored and honey samples, such (Figures 5 and 6), and these samples were as Alternanthera , Byrsonima , Cecropia , dominated by the pollen types Byrsonima , Heteropterys , P. Ju li fl or a ,andSolanum ; howev- B. verticillata , Cecropia , Schefflera er, in a few cases, the order of abundance of these 426 J.S. de Novais et al.

Predominant Pollen Predominant Pollen Predominant Pollen Predominant Pollen Cecropia,, Prosopis juliflora Gomphrena demissa ,, Prosopis juliflora Byrsonima ,, Cecropia Clidemia hirta ,Borreria verticillata,,, Byrsonima Cecropia Schinus, Solanum Schinus , Solanum Solanume Warszewiczia coccinea Chamaecrista , Diodia

Secondary Pollen Secondary Pollen Secondary Pollen Secondary Pollen Heteropterys ,, Myrcia Prosopis Brosimum, Caesalpinioideae type, Byrsonima,, Cecropia Clathrothropis Anacardium occidentale , Borreria juliflora ,, Schinus Solanum , Euphorbia ,, Heteropterys Prosopis nitida ,,,, Clidemia hirta Microtea Myrcia verticillata ,, Byrsonima Emilia fosbergii , Syagrus coronata juliflora ,, Schinus Senna macranthera ,Protium ,, Senna Solanum ,Tapirira Eriope ,, Microtea Tapirira guianensis , Solanum guianensis , Warszewiczia coccinea Warszewiczia coccinea

0 0

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J J M

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M O J J

M S N

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Figure 4. Phenogram generated from the cluster analysis, using the Dice similarity coefficient, of the pollen spectra of 57 samples of honey produced by Tetragonisca angustula (Apidae, Meliponini) in areas of the Lower Amazon (Belterra and Santarém, state of Pará) and of the semiarid region (Itaberaba and Ruy Barbosa, state of Bahia) in Brazil. The main clusters obtained are emphasized (letters A –F ) as well as the dominant pollen types (>45 %) and accessory pollen types (16–45 %) characteristic of the groups. Cophenetic coefficient: 0.9378. genera changed. A similar trend was also ob- availability of food in the foraging radius of served by Imperatriz-Fonseca et al. (1984), who the meliponines, determining the extension of analyzed the food provisions (honey and pollen) their trophic niche. of T. angustula in the state of São Paulo, southeast The cluster analysis here performed did region of Brazil, and by Sosa-Nájera et al. (1994), not allow the establishment of well-defined who conducted research with T. angustula in the groups based only on the climatic data from south of Mexico. the study sites. However, assuming that the According to our studies and to data from beginning, duration, and synchronization of the literature previously cited, some meliponine the flowering process are related, although species prefer certain pollen types for certain not exclusively, to climatic factors (Opler periods of time. However, additional studies et al. 1976; Murphy and Lugo 1986), it is are required to evaluate if this is a real possible to presume that there is a direct preference or simply arises from bees relationship between climatic conditions and exploiting seasonally available pollen. In the pollen spectra composition. case of T. angustula ,Obregónetal.(2013) In arid and semiarid ecosystems, the relation- noted that the workers of a same colony usu- ship between flowering and precipitation is strik- ally tend to be consistent, visiting the same ing because, for most species, the flower buds species and showing preferences for certain generally start blossoming after the rains nectar sources, mainly for species that mass (Machado et al. 1997), changing the physiogno- flowers. These preferences are related to the my of the caatingas . For tropical regions, Opler Melissopalynology of Amazon and caatinga 427

Predominant

p Predominant Pollen Predominant Predominant Pollen Predominant Predominant Pollen Pollen Predomi-

l l Brosimum, Pollen Prosopis juliflora, Pollen Byrsonima , Cecropia Cecropia, nant

P a Senna macranthera, Brosimum, Schinus, Cecropia, Byrsonima, Pollen

n Solanum, Waltheria Solanum n Secondary Pollen i Prosopis juliflora Eriope Clidemia hirta, Clidemia

n n Alternanthera ,, Borreria verticillata Vismia guianensis hirta,

m Secondary Pollen Secondary Pollen Secondary Pollen m Secondary Brosimum ,,, Byrsonima Cecropia Davilla

o a Byrsonima ,, Heteropterys Brosimum, Brosimum ,, Heteropterys Pollen Lindackeria paludosa , Schefflera Secondary kunthii

r r Prosopis juliflora ,, Schinus Prosopis juliflora, Prosopis juliflora , Schinus Acalypha morototoni,, Tabebuia Tapirira guianensis Pollen

P C Solanum, Waltheria Solanum , Virola V. guianensis

t t

v t v

r y

e e

J O M

D F

O A J O M

N S

A J

F J

B B

B S B B S S

I B

R R I

R R

R R R

I R B

S B I

I B 1.0 I

0.9

0.8 L K

0.7 M E

0.6 I

i 0.5

S F J H 0.4 B G 0.3 C D 0.2

0.1 A 0.0

Figure 5. Phenogram generated from the cluster analysis, using the Dice similarity coefficient, of the pollen spectra of 46 samples of pollen stored by Tetragonisca angustula (Apidae, Meliponini) in areas of the Lower Amazon (Belterra and Santarém, state of Pará) and of the semiarid region (Itaberaba and Ruy Barbosa, state of Bahia) in Brazil. The 12 clusters obtained are emphasized (letters A –M ) as well as the dominant pollen types (>45 %) and accessory pollen types (16–45 %) characteristic of the groupings. Cophenetic coefficient: 0.9615.

et al. (1976) state that factors such as the reduction sources by different meliponines, although not of water stress and the isolated decrease in tem- necessarily during the same foraging period. perature can break dormancy and, consequently, In the samples from the caatinga ,thefamilies lead to the synchronized anthesis of several plant with predominant pollen types were species. This synchronization favors cross-polli- Amaranthaceae, Anacardiaceae, Fabaceae, nation, providing flower rewards that attract sev- Malvaceae, Moraceae, Solanaceae, and eral pollinators (Opler et al. 1976). Urticaceae. These plant groups are cited in several The botanical families with predominant pollen studies of the bee flora in this semiarid area (Alves types in the samples from the Amazonian region et al. 2006; Borges et al. 2006;Novaisetal.2006; were Dilleniaceae, Fabaceae, Hypericaceae, Santos et al. 2006). Lamiaceae, Malpighiaceae, Melastomataceae, Roubik and Moreno (2013) state that the Myrtaceae, Rubiaceae, Solanaceae, and predominant pollen is often not an indication Urticaceae. Similar results were found by Rech ofthesourceofthenectar.Thisfactis andAbsy(2011a, b) when working with evident when observing the presence of pol- meliponines of the genera Cephalotrigona , len types related to polleniferous species or Partamona , Ptilotrigona , Scaura , genera (such as Brosimum , Cecropia , Tetragonisca , and Trigona along the canal of Clidemia , Solanum etc.) as the predominant the Negro River in the state of Amazonas. These pollen in honey samples. These pollen spec- results indicate the sharing of the same floral tra provide information on the floral species 428 J.S. de Novais et al.

Figure 6. Climatic data obtained by INMET (2013) for the municipalities of Belterra (a ), Santarém (b ), and Itaberaba (c ). A month was considered dry (emphasized in gray in the graphs) when the total precipitation (mm) was equal to or lower than twice the temperature (°C), according to Bagnouls and Gaussen (1962). that bees visit, but not necessarily the species The pollen spectra of the honey samples re- that they collect nectar from. vealed a higher number of pollen types than the Melissopalynology of Amazon and caatinga 429 pollen spectra of the pollen stored samples. This pollen types than honeys from dry areas. This fact finding possibly reflects the higher number of could be related to differences in the nectar plants that T. angustula needs to visit to find composition at the studied sites, mainly guided nectar in both the Amazonian and the caatinga by climatic factors. In Costa Rica, Biesmeijer areas. Also, the extra plant species found in the et al. (1999) observed that species of Melipona honey samples could come from contamination collected richer nectar at a dry climate site than at after visiting nectar’s flowers. In contrast, the a humid climate site. However, other factors can protein need was supplied by a smaller array of also influence the foraging behavior of bees, such plant species, although this pattern does not nec- as specific need of colonies or interspecific com- essarily indicate a lower collection effort. The petition. Further studies are required to clarify bees may have reduced foraging to a few species these points. but visited a large number of flowers of those Based on the cluster analysis, we conclude the species in search of pollen. pollen data of the honey samples were more con- The cluster analysis performed with honeys sistent in the grouping of samples analyzed per provided two distinct clusters based on the geo- geographic region than the pollen data of the graphical origin of the samples. Results obtained pollen stored samples. The climatic data did not from pollen stored presented lower similarity than allow a consistent grouping of the samples by that provided by honeys. We hypothesize the study sites. more homogeneous is the collection, the higher the similarity between samples. Ecological indices previously calculated by Novais and Absy (2013) ACKNOWLEDGMENTS and Novais et al. (2013, 2014)fromthesamedata set discussed here showed higher values for even- We are extremely grateful to Carleandro Dias and ness in honey samples than pollen stored. The Geancarlo Gouveia for authorizing the collection of more heterogeneous pattern of foraging recorded samples within their private meliponaries. We are also grateful to the regional management of the Company for by pollen stored analysis is due to the concentrat- Agricultural Development of the State of Bahia ed harvest of pollen from certain plant species by (EBDA) for providing logistic support during the sam- T. angustula . On the other hand, the nectar har- plings in Bahia, to the direction of the Mãe Jovina vest revealed low specificity and a more homoge- School of the Agricultural Family for allowing access neous pattern of foraging, resulting in high simi- to the meliponary in Ruy Barbosa, to the Municipal larity between samples. Secretariat of Education of Santarém for authorizing The pattern observed in the cluster analy- the collections within the School of the Forest, to Bruno sis should not be considered as a rule, how- Henrique Andrade-Silva for contributing to the collec- ever. In fact, all foraging strategies are sen- tions and to the processing of the samples, and to sitive to increased competitive pressure Leilton Damascena for providing the map shown here. (Biesmeijer and Slaa 2006), and we have We thank the Brazilian National Council for Scientific and Technological Development (CNPq) for partial not examined this point here. Brazil tends funding of this research and for the scholarships granted to present a gradient in number of species to the authors of this study (processes 575747/2008-0, of plants and stingless bees from semiarid to 143084/2009-7, 303557/2010-9, and 477127/2011-8). Amazon forest areas, i.e., from dry to wet regions. The honey- and pollen-based clusters showed this when group samples were compared from Itaberaba/Ruy Barbosa and Comparaison des spectres de pollens récoltés par Santarém/Belterra, albeit with different simi- Tetragonisca angustula (Apidae, Meliponini) dans la larity levels. basse vallée de l'Amazone (Nord du Brésil) et dans la Caatinga (Nord-Est du Brésil) We found a slight difference in the number of pollen types recorded from Amazonian and analyse de cluster / indice de similarité de Dice / caatinga sites, especially regarding honey entomopalynologie / miel / mélissopalynologie / pollen samples. Honeys from Amazon presented more stocké / abeille sans aiguillon 430 J.S. de Novais et al.

Ein Vergleich der Pollenspektren von Tetragonisca Pesquisa [online] http://www.inmet.gov.br/portal/ angustula (Apidae, Meliponini) aus dem unteren Amazo- index.php?r=bdmep/bdmep (accessed on 21 April 13) nas (Nordbrasilien) und der Caatinge (Nordostbrasilien) Louveaux, J., Maurizio, A., Vorwohl, G. (1978) Methods of melissopalynology. Bee World 59 (4), 139–157 Machado, I.C., Barros, L.M., Sampaio, E.V.S.B. (1997) Clusteranalyse / Dice Ähnlichkeit / Entomopalynologie / Phenology of caatinga species at Serra Talhada, PE, Melissopalynologie / stachellose Bienen / Honig/ Northeastern Brazil. Biotropica 29 (1), 57–68 Pollenvorrat Maia-Silva, C., Silva, C.I., Hrncir, M., Queiroz, R.T., Imperatriz-Fonseca, V.L. (2012) Guia de Plantas Visitadas por Abelhas na Caatinga. Fundação Brasil Cidadão, Fortaleza REFERENCES Marques-Souza, A.C. (2010) Ocorrência do pólen de Podocarpus sp. (Podocarpaceae) nas coletas de Alves, R.M.O., Souza, B.A., Carvalho, C.A.L. (2006) Frieseomelitta varia Lepeletier 1836 (Apidae: Espectro polínico de amostras de mel de Melipona Meliponinae) em uma área de Manaus, AM, Brasil. mandacaia Smith. 1863 (Hymenoptera—Apidae). Acta Bot. Bras. 24 ,558–566 – Acta Sci. Biol. Sci. 28 (1), 33 38 Marques-Souza, A.C., Moura, C.O., Nelson, B.W. (1996) Bagnouls, F., Gaussen, H. (1962) Estação seca e índice Pollen collected by Trigona williana (Hymenoptera: xerotérmico. Boletim Geográfico 20 (169), 338–369. Apidae) in Central Amazonia. Rev. Biol. Trop. 44 (2), (translated into Portuguese by R. S. B. Santos) 567–573 Biesmeijer, J.C., Slaa, E.J. (2006) The structure of eusocial Marques-Souza, A.C., Miranda, I.P.A., Moura, C.O., bee assemblages in Brazil. Apidologie 37 ,240–258 Rabelo, A., Barbosa, E.M. (2002) Características morfológicas e bioquímicas do pólen coletado por Biesmeijer, J.C., Smeets, M.J.A.P., Richter, J.A.P., cinco espécies de meliponíneos da Amazônia Central. Sommeijer, M.J. (1999) Nectar foraging by stingless Acta Amazonica 32 ,217–229 bees in Costa Rica: botanical and climatological influ- ences on sugar concentration of nectar collected by McCune, B., Grace, J. (2002) Analysis of Ecological Com- Melipona . Apidologie 30 ,43–55 munities. MjM Software, Gleneden Beach Borges, R.L.B., Lima, L.C.L., Oliveira, P.P., Silva, F.H.M., Murphy, P.G., Lugo, A.E. (1986) Ecology of tropical dry Novais, J.S., Dórea, M.C., Santos, F.A.R. (2006) O pólen forest. Annu. Rev. Ecol. Syst. 17 ,67–88 no mel do semi-árido brasileiro. In: Santos, F.A.R. (ed.) – Novais, J.S., Absy, M.L. (2013) Palynological examination Apium Plantae, pp. 103 118. IMSEAR, Recife of the pollen pots of native stingless bees from the Erdtman, G. (1960) The acetolysis method—a revised de- Lower Amazon region in Pará, Brazil. Palynology scription. Sv. Bot. Tidskr. 54 (4), 561–564 37 (2), 218–230 Fisch, G., Marengo, J.A., Nobre, C.A. (1998) Uma revisão Novais, J.S., Lima, L.C.L., Santos, F.A.R. (2006) Espectro geral sobre o clima da Amazônia. Acta Amazonica polínico de méis de Tetragonisca angustula Latreille, 28 (2), 101–126 1811 coletados na caatinga de Canudos, Bahia, Brasil. Magistra 18 (4), 257–264 Freitas, B.M., Imperatriz-Fonseca, V.L., Medina, L.M., Kleinert, A.M.P., Galetto, L., Nates-Parra, G., Novais, J.S., Absy, M.L., Santos, F.A.R. (2013) Pollen Quezada-Euán, J.J.G. (2009) Diversity, threats and grains in honeys produced by Tetragonisca angustula conservation of native bees in the Neotropics. (Latreille, 1811) (Hymenoptera: Apidae) in tropical Apidologie 40 ,332–346 semi-arid areas of north-eastern Brazil. Arthropod- Plant Interactions 7 (6), 619–632 Hammer, Ø., Harper, D. A. T., Ryan, P. D. (2001) PAST: Palaeontological Statistics software pack- Novais, J.S., Absy, M.L., Santos, F.A.R. (2014) Pollen age for education and data analysis. types collected by Tetragonisca angustula (Hymenop- Palaeontologia Electronica 4 (1) [online] http:// tera: Apidae) in dry vegetation in Northeastern Brazil. palaeo-electronica.org/2001_1/past/issue1_01.htm Eur. J. Entomol. 111 (1), 25–34 (accessedon17April13) Novais,J.,S.,Absy,M.L.(in press )First IBGE—Instituto Brasileiro de Geografia e Estatística. melissopalynological records in honeys from (2004) Mapa dos biomas brasileiros. IBGE, Rio de Tetragonisca angustula (Latreille, 1811) in Lower Janeiro. [online] http://www.ibge.gov.br (accessed on Amazon, Brazil: pollen spectrum and concentration. 2 May 13) J. Apic. Res. Imperatriz-Fonseca, V.L., Kleinert-Giovannini, A., Obregón, D., Rodríguez, C.Á., Chamorro, F.J., Nates-Parra, Cortopassi-Laurino, M., Ramalho, M. (1984) Hábitos G. (2013) Botanical origin of pot-honey from de coleta de Tetragonisca angustula angustula Tetragonisca angustula Latreille in Colombia. In: Vit, Latreille (Apidae, Meliponinae). Boletim de Zoologia P., Pedro, S.R.M., Roubik, D. (eds.) Pot-Honey: A Leg- da Universidade de São Paulo 8 ,115–131 acy of Stingless Bees, pp. 337–346. Springer, New York INMET—Instituto Nacional de Meteorologia. (2013) Opler, P.A., Frankie, G.W., Baker, H.G. (1976) Rainfall as a Banco de Dados Meteorológicos para Ensino e factor in the release, timing, and synchronization of Melissopalynology of Amazon and caatinga 431

anthesis by tropical trees and shrubs. J. Biogeogr. 3 , Roubik, D., Moreno, P.J.E. (2013) How to be a bee-botanist 231–236 using pollen spectra. In: Vit, P., Pedro, S.R.M., Roubik, D. (eds.) Pot-honey: a legacy of stingless bees, pp. Prado, D. (2003) As caatingas da América do Sul. In: Leal, – I.R., Tabarelli, M., Silva, J.M.C. (eds.) Ecologia e 295 314. Springer, New York conservação da caatinga, pp. 3–73. Editora Santana, A.L.A., Fonseca, A.A.O., Alves, R.M.O., Universitária da UFPE, Recife Carvalho, C.A.L., Melo, P.A., Silva, E.S., Souza, B.A., Jesus, J.N., Sodré, G.S. (2011) Tipos polínicos Ramalho, M., Imperatriz-Fonseca, V.L., Kleinert- em amostras de méis de abelhas sem ferrão de Giovannini, A., Cortopassi-Laurino, M. (1985) Exploi- municípios do semiárido baiano. Magistra 23 (3), tation of floral resources by Plebeia remota Holmberg 134–139 (Apidae, Meliponinae). Apidologie 16 (3), 307–330 Santos, F.A.R., Oliveira, J.M., Oliveira, P.P., Leite, K.R.B., Rech, A.R., Absy, M.L. (2011a) Pollen sources used by Carneiro, C.E. (2006) Plantas do semi-árido species of Meliponini (Hymenoptera: Apidae) along importantes para as abelhas. In: Santos, F.A.R. (ed.) the Rio Negro channel in Amazonas, Brazil. Grana Apium Plantae, pp. 61–86. IMSEAR, Recife 50 (2), 150–161 Sosa-Nájera, M.S., Martínez-Henández, E., Lozano- Rech, A.R., Absy, M.L. (2011b) Pollen storages in nests of García, M.S., Cuadriello-Aguiar, J.I. (1994) bees of the genera Partamona , Scaura and Trigona Nectaropolliniferous sources used by Trigona (Hymenoptera, Apidae). Rev. Bras. Entomol. 55 (3), (Tetragonisca ) angustula in Chiapas, southern Méxi- 361–372 co. Grana 33 (4), 225–230